Tissue separation using QuPath

Hi everyone,

I am trying to do tissue separation using QuPath - I tried using pixel classification, and although it was successful in identifying tissue such as muscle, it highlighted areas within other regions. Furthermore, if I tried to separate between tissue regions such as muscularis mucosae and muscularis externa, the classifier would highlight all the muscle regardless of the location. Attaching picture below of example tissue of how I would like the tissue to annotate the regions:

I was suggested using object classifier but it requires information from “Cell detection” function which is not useful in this case (at least in my opinion).

Would anyone have any suggestions on how to go about this?

Thanks.

FYI - the staining is of Picro-Sirius Red for collagen.

The pixel classifier is not magic, but if you have enough stain and or structure variation, and enough well varied training data, it can do a lot. That looks doable, but that’s rather easier to say having not tried your particular example.

If it is not working, even with using stain intensity instead of RGB and while using texture features, you may want to look into classifying objects/SLICs, which can have additional measurement types beyond what the pixel classifier can use (Haralick, distance tiles, smoothing, etc).